title: [Effects of pregnancy on the ROS, NO, cytokine levels and lymphocytes activation from mouse peripheral blood].
abstract: OBJECTIVE
  To investigate the influence of pregnancy on the production of reactive oxygen species (ROS) from mouse peripheral blood neutrophils (PMN), the levels of NO and cytokines from serum, the activation of T lymphocytes, and initially find the immune regulation effects of pregnancy on the mouse peripheral blood lymphocytes.
  
  
  METHODS
  Take the BALB/c mice which were at the mid trimester of pregnancy (day 14) as the object, full blood staining using ROS probe 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) combing with flow cytometry was used to test the levels of ROS from PMN. The production of NO from peripheral blood serum were analyzed by Griess kit while the soluble cytokines interleukin (IL) 6, IL-10, monocyte chemotactic protein 1 (MCP-1), interferon γ (IFN-γ), tumor necrosis factor α (TNF-α) and IL-12 were detected by liquid protein quantitative technology cytometric bead array (CBA) using flow cytometry. The activation of peripheral blood lymphocytes at early, middle and later phases which marked with CD69, CD25 and CD71 respectively were tested by flow cytometry and two-color fluorescent staining.
  
  
  RESULTS
  Comparing to the normal non-pregnant mouse, pregnancy obviously promoted the production of ROS from PMN (101.1±2.2 versus 134.5±10.3, P<0.05). Comparing to the normal non-pregnant mouse, pregnancy obviously promoted the secretion of NO [(22.7±0.7) versus (36.3±1.2) µmol/L, P<0.01]. In normal non-pregnant mouse, the serum levels of IL-6, IL-10, MCP-1, IFN-γ, TNF-α and IL-12 were (9.3±0.5), (26.7±0.9), (21.2±1.6), (14.5±1.8), (22.6±1.6) and (8.4±1.2) pg/ml, while in pregnancy group the levels were (26.5±1.0), (40.4±2.5), (25.1±0.7), (457.4±17.9), (93.2±4.3) and (7.5±0.9) pg/ml correspondingly; the levels of IL-6, IFN-γ, TNF-α from peripheral blood serum (P<0.01), while had no effects on the production of IL-10 and MCP-1 (P>0.05). About the CD+3 T lymphocytes activation, in normal non-pregnant mouse, the CD69, CD25 and CD71 expression rate were (0.43±0.15)%, (5.13±0.25)% and (0.37±0.11)%, while in pregnancy group the CD69, CD25 and CD71 expression rate were (0.40±0.10)%, (6.17±0.40)% and (6.10±0.31)%. The levels of middle and later phases markers as CD25 and CD71 were highly up-regulated (P<0.05), while the early phase action CD69 had no obvious variation (P>0.05).
  
  
  CONCLUSIONS
  The mid trimester of pregnancy promoted the production of ROS from PMN, the levels of NO, IL-6, IFN-γ, TNF-α from peripheral blood serum, and the middle- and later-phase activation of T lymphocytes.
authors.affiliation: Department of Obstetrics and Gynecology, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China.
authors.forename: Fang
authors.initials: F
authors.lastname: He
authors.forename: Dun-jin
authors.initials: DJ
authors.lastname: Chen
authors.forename: Lian-ying
authors.initials: LY
authors.lastname: Li
authors.forename: Qi-xian
authors.initials: QX
authors.lastname: Chen
mesh_terms: D000818:Animals; D016207:Cytokines / Q000097:blood*; D005260:Female; D008213:Lymphocyte Activation; D051379:Mice; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D009569:Nitric Oxide / Q000097:blood*; D011247:Pregnancy / Q000097:blood / Q000276:immunology*; D011262:Pregnancy Trimester, Second; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D017382:Reactive Oxygen Species; D009569:Nitric Oxide